Difference between revisions of "Part:BBa K5034201"
 
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===Sequence and Features===
 
===Sequence and Features===
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<partinfo>BBa_K5034201 SequenceAndFeatures</partinfo>
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This backbone consists of several parts, including promoters, terminators, resistance genes, and prefix and suffix we added.
  
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===Experimental Characterization and results===
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<img style="width:50%;height:auto;" src="https://static.igem.wiki/teams/5034/engineering/pbbr1mcs-terminator.png">
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Figure 1: Plasmid diagram of backbone
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<partinfo>BBa_K5034201 SequenceAndFeatures</partinfo>
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===Origin===
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It's antibiotic-resistant derivatives of the broad-host-range (bhr) cloning vector.We get the pBBR1MCS-2 plasmid on the addgene, and add a terminator rrnBT1-T7Te(BBa-B0015) to it.
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===Experimental Characterization and results===
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We performed double enzyme digestion on the plasmid and obtained a fragment of approximately 5000bp(Fig.2)
 
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<div align="center">
 
<div align="center">
 
<img style="width:50%;height:auto;" src="https://static.igem.wiki/teams/5034/results/fig23.jpg">
 
<img style="width:50%;height:auto;" src="https://static.igem.wiki/teams/5034/results/fig23.jpg">
 
<p>
 
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Figure 1: Fragment obtained by double enzyme digestion of plasmid backbone
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Figure 2: Fragment obtained by double enzyme digestion of plasmid backbone
 
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We then introduce it to <i>S.oneidensis</i>.
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===Applications===
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===Chassis and Genetic Context===
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We successfully expressed in ''Shewanella onediensis'' MR-1.
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===Potential Applications===
  
 
1. It can be used in microbial fuel cells to study and enhance the extracellular electron transfer capabilities such as in <i>S. oneidensis</i>.
 
1. It can be used in microbial fuel cells to study and enhance the extracellular electron transfer capabilities such as in <i>S. oneidensis</i>.
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2. It can be used as a backbone for CRISPR-based genome editing systems, enabling precise genetic modifications.
 
2. It can be used as a backbone for CRISPR-based genome editing systems, enabling precise genetic modifications.
  
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3. It was used in our project to carry components related to phosphorus metabolism. Our results indicate that this component is a good backbone for carrying phosphorus metabolism related components. So it may be able to be used to solve the problems of regulating the electricity production and phosphorus accumulation of <i>S.oneidensis</i>. Thus providing the possibility for organisms to generate electricity while removing pollution.
 
===Reference===
 
===Reference===
  
 
<i>1. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM 2nd, Peterson KM. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene. 1995 Dec 1;166(1):175-6. doi: 10.1016/0378-1119(95)00584-1. PMID: 8529885.</i>
 
<i>1. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM 2nd, Peterson KM. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene. 1995 Dec 1;166(1):175-6. doi: 10.1016/0378-1119(95)00584-1. PMID: 8529885.</i>

Latest revision as of 07:46, 2 October 2024


pBBR1MCS-terminator plasmid

Description

The pBBR1MCS plasmid is a synthetic plasmid backbone used in various genetic engineering applications in microbial systems. We modified the original pBBR1MCS and added a double terminator rrnBT1-T7Te(BBa_B0015) downstream the Biobrick suffix.

Sequence and Features

This backbone consists of several parts, including promoters, terminators, resistance genes, and prefix and suffix we added.

Figure 1: Plasmid diagram of backbone


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4981
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2834
    Illegal NotI site found at 4987
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4981
    Illegal BglII site found at 3580
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 4981
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 4981
    Plasmid lacks a suffix.
    Illegal XbaI site found at 4996
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 562
    Illegal NgoMIV site found at 4244
    Illegal NgoMIV site found at 4527
    Illegal AgeI site found at 402
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.

Origin

It's antibiotic-resistant derivatives of the broad-host-range (bhr) cloning vector.We get the pBBR1MCS-2 plasmid on the addgene, and add a terminator rrnBT1-T7Te(BBa-B0015) to it.

Experimental Characterization and results

We performed double enzyme digestion on the plasmid and obtained a fragment of approximately 5000bp(Fig.2)

Figure 2: Fragment obtained by double enzyme digestion of plasmid backbone

We then introduce it to S.oneidensis.

Chassis and Genetic Context

We successfully expressed in Shewanella onediensis MR-1.

Potential Applications

1. It can be used in microbial fuel cells to study and enhance the extracellular electron transfer capabilities such as in S. oneidensis.

2. It can be used as a backbone for CRISPR-based genome editing systems, enabling precise genetic modifications.

3. It was used in our project to carry components related to phosphorus metabolism. Our results indicate that this component is a good backbone for carrying phosphorus metabolism related components. So it may be able to be used to solve the problems of regulating the electricity production and phosphorus accumulation of S.oneidensis. Thus providing the possibility for organisms to generate electricity while removing pollution.

Reference

1. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM 2nd, Peterson KM. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene. 1995 Dec 1;166(1):175-6. doi: 10.1016/0378-1119(95)00584-1. PMID: 8529885.